TruSeq Stranded Total RNA

A robust, highly scalable whole-transcriptome analysis (RNA-Seq) solution for a variety of species and sample types, including human, mouse, and formalin-fixed, paraffin-embedded (FFPE) tissue.

11.5 hr

Assay time

5.5 hr

Hands-on time

0.1 – 1 ug high-quality total RNA. Lower-quality samples may require further optimization.

Input quantity

See full details in the specifications table

The products previously known as TruSeq Stranded Total RNA Library Prep Kit with Ribo-Zero Human/Mouse/Rat (Cat. No. RS-122-2201, RS-122-2202, and RS-122-2203) and TruSeq Stranded Total RNA Library Prep Kit with Ribo-Zero Gold (Cat. No. RS-122-2301, RS-122-2302, and RS-122-2303) are now called TruSeq Stranded Total RNA Human/Mouse/Rat and TruSeq Stranded Total RNA Gold, respectively. The product configuration has changed. You'll need to purchase the library prep and index adapter products separately (instead of in a kit).

Overview

TruSeq Stranded Total RNA provides a clear view of the transcriptome with a streamlined, cost-efficient, and scalable solution for total RNA analysis with sequencing.

Whole-Transcriptome analysis with precise measurement of strand orientation

Compatible with a wide range of samples, including low-quality DNA/RNA and formalin-fixed, paraffin-embedded (FFPE) samples, TruSeq Stranded Total RNA couples all of the benefits of TruSeq RNA technology with Ribo-Zero ribosomal RNA reduction chemistry. The product enables analysis of coding and multiple forms of noncoding RNA with precise measurement of strand orientation, uniform coverage, and high-confidence discovery of features such as alternative transcripts, gene fusions, and allele-specific expression.

Species compatibility

TruSeq Stranded Total RNA is designed for human, mouse and rat sequences, but may be applicable to a variety of eukaryotic species.

Flexible solutions for a range of study designs

TruSeq Stranded Total RNA is available in multiple versions to fit specific study design needs:

  • TruSeq Stranded Total RNA Human/Mouse/Rat removes cytoplasmic rRNA

  • TruSeq Stranded Total RNA Gold removes both cytoplasmic and mitochondrial rRNA

  • The total RNA kits are also compatible with the new unique dual (UD) index kits, IDT for Illumina - TruSeq RNA UD Indexes, which allows multiplexing of up to 96 samples

Fast, automation-friendly workflow

Prepare libraries for Illumina sequencing in less than a day. Illumina Qualified Methods are available on a range of automation platforms through our partners.


Specifications


Required Products

Illumina now offers modular product ordering to enable flexibility in your workflows. You may order any of the following library prep products with any of the following indexes. 
/ Results

Applications

Example workflow


Project recommendations

Instrument Recommended number of samples Read length
NextSeq 550 System

RNA profiling: 6-20 samples per run (based on 20 million reads per sample)
Transcriptome analysis: 2-8 samples per run (based on 50 million reads per sample)

2 x 75 bp

NovaSeq 6000 System

RNA profiling (samples per run, dual flow cell): S1: 160, S2: 320, S4: 768. Limited by index combinations (dual). Based on 20 million reads.
Transcriptome analysis (samples per run, dual flow cell): S1: 64, S2: 128, S4: 400. Limited by index combinations (dual). Based on 50 million reads. 

≤ 2 × 100 bp


Related applications and methods

Compare

TruSeq Stranded Total RNA Illumina Stranded Total RNA Prep with Ribo-Zero Plus or Ribo-Zero Plus Microbiome
Assay time 11.5 hr ~7 hr
Automation capability Liquid handling robots Liquid handling robots
Automation details Explore available automation methods
Content specifications Captures coding RNA plus multiple forms of non-coding RNA Captures coding RNA plus multiple forms of non-coding RNA
Description For whole-transcriptome sequencing studies. The Total RNA version depletes cytoplasmic rRNA while the Total RNA Gold version depletes cytoplasmic and mitochondrial rRNA.

For the preparation of whole-transcriptome sequencing-ready libraries from low RNA inputs and low-quality samples that supports a wide range of RNA-Seq applications. Includes Ribo-Zero Plus for single-tube depletion of rRNA and globin RNA from multiple species to facilitate rich transcriptome analyses. 

Hands-on time 5.5 hr < 3 hr
Input quantity 0.1 – 1 ug high-quality total RNA. Lower-quality samples may require further optimization.

1–1000 ng total RNA for standard-quality RNA samples. 10 ng total RNA minimum recommended for optimal performance and FFPE samples 

Instruments NextSeq 550 System, NextSeq 2000 System, NextSeq 1000 System, NovaSeq X System, NextSeq 500 System, NovaSeq 6000 System, NovaSeq X Plus System NextSeq 550 System, NextSeq 2000 System, NextSeq 1000 System, NovaSeq X System, NextSeq 500 System, NovaSeq 6000 System, NovaSeq X Plus System
Mechanism of action Bead-based rRNA depletion, cDNA synthesis, and PCR Enzymatic rRNA depletion, ligation-based addition of adapters and indexes
Method Whole-transcriptome sequencing Whole-transcriptome sequencing, Metatranscriptome sequencing
Multiplexing Low-throughput kits: Pool up to 12 samples. Or pool up to 24 samples with sets A and B together, High-throughput kits: Prepare 96 indexed samples using either combinatorial dual indexes or unique dual indexes Up to 384 Unique Dual Indexes (UDIs)
Nucleic acid type RNA RNA
Specialized sample types FFPE Tissue Microbiome Samples, Low-Input Samples, FFPE Tissue
Species category Mouse, Human, Rat Mouse, Human, Rat, Bacteria
Species details rRNA Removal: Depletes human, mouse, and rat rRNA

Ribo-Zero Plus depletes abundant transcripts from multiple species including: human cytoplasmic and mitochondrial rRNA, mouse rRNA, rat rRNA, bacteria (E. coli and B. subtilis) rRNA, and human beta globin transcripts 

Ribo-Zero Plus Microbiome depletes abundant transcripts from multiple species including: those depleted in Ribo-Zero Plus as well as diverse microbial species and ATCC MSA-2002, MSA-2005, and MSA-2006 

Technology Sequencing Sequencing
Variant class Single nucleotide polymorphisms (SNPs), Gene fusions, Novel transcripts, Transcript variants Single nucleotide polymorphisms (SNPs), Gene fusions, Novel transcripts, Transcript variants

Selection tools:

Figures

High quality strand information

Consistent, precise measurement of RNA abundance is reflected by high reproducibility between technical replicates. These technical replicates of FFPE tissue show high concordance, indicating robust library prep performance. Axes are log2(FPKM). R2 value is shown.

Even coverage across transcripts

TruSeq Stranded Total RNA gives excellent coverage across the top 1000 expressed transcripts in both fresh-frozen (FF, top) and FFPE (bottom) tumor and matched normal breast tissue, with > 98% aligned stranded reads. X-axis: position along transcript, Y-axis: percent coverge of combined reads.

Library prep (4)

TruSeq® Stranded Total RNA Library Prep Human/Mouse/Rat (48 Samples)

20020596

Includes RiboZero beads to deplete rRNA from human, mouse, and rat, PCR master mix to transcript RNA into cDNA , and reagents for preparing 48 samples. Purchase index adapters and Purification Beads separately.

Sign in to add to cart or see pricing.

List Price:

Discounts:

TruSeq® Stranded Total RNA Library Prep Human/Mouse/Rat (96 Samples)

20020597

Includes RiboZero beads to deplete rRNA from human, mouse, and rat, PCR master mix to transcript RNA into cDNA , and reagents for preparing 48 samples. Purchase index adapters and Purification Beads separately.

Sign in to add to cart or see pricing.

List Price:

Discounts:

TruSeq Stranded Total RNA Library Prep Gold (48 Samples)

20020598

Includes RiboZero beads to deplete rRNA and mitochondrial rRNA from human, mouse, and rat, PCR master mix to transcript RNA into cDNA, and reagents for preparing 48 samples. Purchase index adapters separately. Purchase index adapters and Purification Beads separately.

Sign in to add to cart or see pricing.

List Price:

Discounts:

TruSeq® Stranded Total RNA Library Prep Gold (96 Samples)

20020599

Includes RiboZero beads to deplete rRNA and mitochondrial rRNA from human, mouse, and rat, PCR master mix to transcript RNA into cDNA, and reagents for preparing 48 samples. Purchase index adapters separately. Purchase index adapters and Purification Beads separately.

Sign in to add to cart or see pricing.

List Price:

Discounts:

Index adapters (4)

TruSeq RNA CD Index Plate (96 Indexes, 96 Samples)

20019792

TruSeq Stranded RNA HT Kit 96 Samples Adapter Plate Box

Sign in to add to cart or see pricing.

Request pricing

List Price:

Discounts:

TruSeq RNA Single Indexes Set A (12 Indexes, 48 Samples)

20020492

Includes 12 of 24 indexes (Indexes: 2, 4, 5, 6, 7, 12, 13, 14, 15, 16, 18, 19) for preparing 48 individual RNA samples.

Sign in to add to cart or see pricing.

List Price:

Discounts:

TruSeq RNA Single Indexes Set B (12 Indexes, 48 Samples)

20020493

Includes 12 of 24 indexes (Indexes: 1, 3, 8, 9, 10, 11, 20, 21, 22, 23, 25, 27) for preparing 48 individual RNA samples.

Sign in to add to cart or see pricing.

List Price:

Discounts:

IDT for Illumina – TruSeq RNA UD Indexes v2 (96 Indexes, 96 Samples)

20040871

Includes 96, 8 bp indexes sufficient for labeling 96 samples and is compatible with TruSeq Stranded mRNA and TruSeq Stranded Total RNA Library Prep Kits. Version 2 contains improvements for 8 of the 192 index oligos in plate 20022371, with no changes to the remaining 184. Purchase library preps and probe panels separately.

Sign in to add to cart or see pricing.

List Price:

Discounts:

Accessory products (1)

Illumina Ribo-Zero plus rRNA Depletion Kit (96 Samples)

20037135

Includes Ribo-Zero Plus depletion module for depletion of ribosomal RNA from human, mouse, rat, and bacteria and globin transcripts for 96 samples. Purchase purification beads, library prep reagents and index adapters separately. For complete kit, including library prep reagents, order the Illumina Stranded Total RNA with Ribo-Zero Plus kit.

Sign in to add to cart or see pricing.

List Price:

Discounts:

Services (1)

TruSeq Total RNA Training

15044069

Sign in to add to cart or see pricing.

Request pricing

List Price:

Discounts:

Showing of

Selection summary

Product

Qty

Unit Price

Order the NovaSeq X Series

Advanced chemistry, optics, and informatics combine to deliver exceptional speed and data quality, outstanding throughput and scalability.

MiSeq Reagent Kit v3

Optimized reagent kit that provides increased cluster density and read length, improving sequencing quality scores compared to earlier versions.

TruSight Oncology 500

Enable CGP with a large pan-cancer panel covering all major variant classes plus gene signatures (TMB, MSI, and HRD) from FFPE tissue.

Illumina DNA Prep

A fast, integrated workflow for preparing libraries for use in a wide range of sequencing applications.

Limited Use Label License: This product and its use are the subject of one or more issued and/or pending U.S. and foreign patent applications owned by Max Planck Gesellschaft, exclusively licensed to New England Biolabs, Inc. and sublicensed to Illumina, Inc. The purchase of this product from Illumina, Inc., its affiliates, or its authorized resellers and distributors conveys to the buyer the non-transferable right to use the purchased amount of the product and components of the product by the buyer (whether the buyer is an academic or for profit entity). The purchase of this product does not convey a license under any claims in the foregoing patents or patent applications direct to producing the product. The buyer cannot sell or otherwise transfer this product or its components to a third party or otherwise use the product for the following COMMERICAL PURPOSES: (I) use of the product or its components in manufacturing; or (2) use of the product or its components for therapeutic or prophylactic purposes in human or animals.

Speak with a specialist

Reach out for information about our products and services, or get answers to questions about our technology.